CN103130904A - High-valued utilization method for patinopecten yessoensis offal - Google Patents
High-valued utilization method for patinopecten yessoensis offal Download PDFInfo
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Abstract
The invention relates to a high-valued utilization method for patinopecten yessoensis offal. The high-valued utilization method for the patinopecten yessoensis offal is technically characterized in that the patinopecten yessoensis offal is cleanly washed, weighted, added with water and made to homogenate. Enzymolysis is carried out on the homogenate by adding trypsin and bacillus subtilis protease in the homogenate. Enzymatic hydrolysate is centrifuged, and supernate and precipitate are separated. The supernate is added with hydrogen peroxide so as to decolor the supernate, the supernate is centrifuged, the potential of hydrogen (PH) is adjusted, acidic and neutral protein is eliminated through centrifugation, and alcohol precipitation is carried out on the supernate, so that a crude product of crude glycosaminoglycan is obtained, and the yield rate is 0.265%. The crude product is dissolved by being added with water, 0.4% trypsin are added to carry out enzymolysis on the crude product for a second time, then the crude product is centrifuged, alcohol precipitation is carried out on the supernate, cetyl trimethyl ammonium bromide (CTAB) is added, and the supernate is centrifuged. The precipitate is dissolved in a sodium acetate solution, dissociation liquid is obtained and dialyzed, and alcohol precipitation is carried out on the dissociation liquid, so that highly purified glycosaminoglycan is obtained. The purity of the glycosaminoglycan can reach more than 90%. The glycosaminoglycan has good antioxygenation and can be used as a raw material of functional foods. All protein precipitate is mixed and is used as a good protein source for feeding animals. The high-valued utilization method for the patinopecten yessoensis offal has advantages that the offal of processed scallops adductor of patinopecten yessoensis is comprehensively utilized in a high-valued mode, and non-pollution and zero emission are achieved in the whole technological process.
Description
Technical field
The invention belongs to food technology field, relate to the tankage higher value application, especially a kind of method of Patinopecten yessoensis tankage higher value application.
Background technology
It is northern that Patinopecten yessoensis originates in Japan, waters, Russian Thousand Islands south, Hokkaido, Japan and Honshu, be Large Scale Cold water-based shellfish, now having introduced China, and propagated, bred production artificially at Northern Coastal Regions such as Shandong, Liaoning, is the important kind of shellfish fishery products in recent years.Patinopecten yessoensis contains abundant unsaturated fatty acids EPA and DHA, also has the effects such as enriching yin, kidney tonifying, and the diseases such as in poor health, poor appetite, malnutrition are had good curative effect.The closed shell flesh of Patinopecten yessoensis is thick, and the meat flavour deliciousness is nutritious, is widely used in to make shellfish post, the needs of satisfying the market.The tankage utilization ratio of processing shellfish post is low at present, causes the waste of resource.The present invention chooses the Patinopecten yessoensis tankage, extracts high-purity glycosaminoglycan, for cosmetic industry, health products trade provide high-quality raw material, prepares simultaneously the high-quality animal proteinum, satisfies the needs of animal cultivation industry.
By retrieval, find following three pieces of publication documents relevant to present patent application: 1, process for extracting comb shell polysaccharide (ZL200510047410.2).The invention provides a kind of extracting method of comb shell polysaccharide, described method comprises raw material processing, homogenate, ultrasonic, enzymolysis, separation, concentrated, alcohol precipitation, dry processing step.The extraction preparation method (CN1746191) of 2, a kind of active substance with vascular protection effect---comb scallop glycosaminoglycan, its extracting and preparing technique is as follows: the chlamys farreri visceral mass is weighed, add water and make homogenate, add proteolytic enzyme in homogenate, reacting by heating gets enzymolysis solution; Enzymolysis solution is centrifugal in whizzer, discard throw out, centrifugate is added the secondary ethanolic soln, form precipitation; Throw out adding distil water after separating is made into aqueous solution post-heating and adds gac, cooling by centrifugal, collect supernatant liquor, to the supernatant liquor suction filtration, get clear liquid; With clear liquid filter membrane ultrafiltration, get the filtrate that molecular weight is 5k～100k; The filtrate lyophilize is namely made comb scallop glycosaminoglycan.The method is simple to operate, reaction conditions is gentle, with low cost, yield and purity high, also can reclaim in a large number protein when extracting glycosaminoglycan, be conducive to the comprehensive utilization of chlamys farreri visceral mass, the material of extraction has important value for control cardiovascular disease.3, a kind of extracting method of bay scallop polysaccharide (CN201010552250.8), present method adopts following steps: with bay scallop tissue mashing post-drying or freeze-drying, soak to reflux through reagent and take off oligosaccharides, pigment; Carry out lixiviate take water as solvent; The centrifugal supernatant liquor that gets; Rotary evaporation in vacuo is concentrated; Add dehydrated alcohol in concentrated solution, ethanol reaches finite concentration makes polysaccharide precipitation; Be drying to obtain the bay scallop polysaccharide.
By comparison, the technical scheme of above-mentioned publication document and extraction object all have different (secondary enzymolysis, the CTAB complexing purifying) of essence from present patent application, and present patent application has novelty.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art part, a kind of method of Patinopecten yessoensis tankage higher value application is provided, sufficient raw, the extraction cost of the method are low, for the widespread use of satisfying cosmetic industry, health products trade and extra large precious animal farming industry provides possibility.
The present invention is achieved through the following technical solutions:
A kind of method of Patinopecten yessoensis tankage higher value application, step is as follows:
⑴ raw material is processed: after the Patinopecten yessoensis tankage are cleaned, add distilled water homogenate, form homogenate;
⑵ enzymolysis: add trypsinase and bacillus subtilis neutral proteinase in above-mentioned homogenate, under 40 ℃ of water bath condition enzymolysis 3.5-4 hour, after enzymolysis solution boils the enzyme that goes out, the centrifugal 10min of 5000r/min, separation of supernatant and precipitation;
⑶ decolouring: add the hydrogen peroxide decolouring in supernatant liquor;
⑷ remove protein: first transfer pH to 2.0, the centrifugal precipitation of removing with HCl; Transfer pH to 7.0, the centrifugal precipitation of removing with NaOH again;
⑸ alcohol deposition method extracts raw sugar amine glycan: supernatant liquor slowly adds ethanol to pure content under constantly stirring be 70%, under 4 ℃ of conditions standing 24 hours, remove supernatant liquor, and reclaim raw sugar amine glycan after the centrifugal 10min of 8000r/min;
⑹ secondary enzymolysis: raw sugar amine glycan is dissolved in distilled water, adds 0.4% trypsinase secondary enzymolysis, pH8.0, enzymolysis time 3 hours boils the centrifugal 10min of 5000r/min after the enzyme that goes out, separation of supernatant and precipitation;
⑺ precipitate merging: all albumen precipitations merge, and make animal protein source;
⑻ CTAB in conjunction with dissociate: supernatant liquor adds the 5%CTAB(cetyl trimethylammonium bromide of 2 times of volumes), under 45 ℃, precipitation is 1-2 hour, the centrifugal 30min of 8000r/min, collecting precipitation, under 45 ℃, precipitation is dissolved in the CH3COONa solution of 2mol/L, the GAG-CTAB complex compound is dissociated, form dissociation solution;
⑼ alcohol precipitation, dialysis, alcohol precipitation again: above-mentioned dissociation solution adds ethanol, make the alcohol amount reach 70%, the centrifugal 10min of 8000r/min, collecting precipitation is dissolved in distilled water with precipitation, dialysis tubing dialysis 48h with the amount of damming 7000, add the Ethanol-Acetic Acid sodium reagent in dialyzate, make the alcohol amount reach 70%, press alcohol deposition method and reclaim glycosaminoglycan, with the throw out oven dry, namely make pure mud snail glycosaminoglycan.
And, in described step ⑴ the add-on of distilled water be tankage weight 2-3 doubly.
And in described step ⑵, the total addition level of trypsinase and bacillus subtilis neutral proteinase is the 0.8-1% of tankage gross weight, and trypsinase and bacillus subtilis neutral proteinase respectively account for half.
And, the addition of hydrogen peroxide is the 5-7% of supernatant liquor weight in described step ⑶.
Advantage of the present invention and positively effect are:
1, present method reaction conditions is gentle, glycosaminoglycan purity is high, extracts with the glycosaminoglycan of other shellfishes and compares, work simplification, time shorten.
2, present method adopts the Patinopecten yessoensis tankage as object, and material cost is low, and added value of product is high, and a whole set of technical process is pollution-free, zero release.
3, present method also can reclaim protein in a large number when extracting glycosaminoglycan, be conducive to the comprehensive utilization of Patinopecten yessoensis tankage, the glycosaminoglycan of extracting has stronger resistance of oxidation, and the protein of recovery can be used as the good protein source of extra large precious animal farming industry.
Embodiment
Below in conjunction with embodiment, the present invention is further described, and following embodiment is illustrative, is not determinate, can not limit protection scope of the present invention with following embodiment.
The object that the present embodiment adopts is to be selected from the Patinopecten yessoensis tankage.
A kind of method of Patinopecten yessoensis tankage higher value application comprises the following steps:
⑴ raw material is processed: after the Patinopecten yessoensis tankage are cleaned, add distilled water homogenate, form homogenate;
⑵ enzymolysis: add trypsinase and bacillus subtilis neutral proteinase in above-mentioned homogenate, under 40 ℃ of water bath condition enzymolysis 3.5-4 hour, after enzymolysis solution boils the enzyme that goes out, the centrifugal 10min of 5000r/min, separation of supernatant and precipitation;
⑶ decolouring: add the hydrogen peroxide decolouring in supernatant liquor;
⑷ remove protein: first transfer pH to 2.0, the centrifugal precipitation of removing with HCl; Transfer pH to 7.0, the centrifugal precipitation of removing with NaOH again;
⑸ alcohol deposition method extracts raw sugar amine glycan: supernatant liquor slowly adds ethanol to pure content under constantly stirring be 70%, under 4 ℃ of conditions standing 24 hours, remove supernatant liquor, and reclaim raw sugar amine glycan after the centrifugal 10min of 8000r/min;
⑹ secondary enzymolysis: raw sugar amine glycan is dissolved in distilled water, adds 0.4% trypsinase secondary enzymolysis, pH8.0, enzymolysis time 3 hours boils the centrifugal 10min of 5000r/min after the enzyme that goes out, separation of supernatant and precipitation;
⑺ precipitate merging: all albumen precipitations merge, and make animal protein source.
⑻ CTAB in conjunction with dissociate: supernatant liquor adds the 5%CTAB(cetyl trimethylammonium bromide of 2 times of volumes), under 45 ℃, precipitation is 1-2 hour, the centrifugal 30min of 8000r/min, collecting precipitation, under 45 ℃, precipitation is dissolved in the CH3COONa solution of 2mol/L, the GAG-CTAB complex compound is dissociated, form dissociation solution;
⑼ alcohol precipitation, dialysis, alcohol precipitation again: above-mentioned dissociation solution adds ethanol, make the alcohol amount reach 70%, the centrifugal 10min of 8000r/min, collecting precipitation is dissolved in distilled water with precipitation, dialysis tubing dialysis 48h with the amount of damming 7000, add the Ethanol-Acetic Acid sodium reagent in dialyzate, make the alcohol amount reach 70%, press alcohol deposition method and reclaim glycosaminoglycan, with the throw out oven dry, namely make pure glycosaminoglycan.
Example 1
⑴ raw material is processed: after the Patinopecten yessoensis tankage are cleaned, add the distilled water homogenate of 2 times of weight, make homogenate;
⑵ enzymolysis: add trypsinase and bacillus subtilis neutral proteinase in above-mentioned homogenate, the enzyme addition is 0.8% of tankage gross weight, and trypsinase and bacillus subtilis neutral proteinase respectively account for half, and enzymolysis is 3.5 hours under 40 ℃ of water bath condition; Separate and get supernatant liquor: after enzymolysis solution boils the enzyme that goes out, the centrifugal 10min of 5000r/min, separation of supernatant and precipitation;
⑶ decolouring: add 5% hydrogen peroxide decolouring in supernatant liquor;
⑷ remove protein: first transfer pH to 2.0, the centrifugal precipitation of removing with HCl; Transfer pH to 7.0, the centrifugal precipitation of removing with NaOH again;
⑸ alcohol deposition method extracts raw sugar amine glycan: supernatant liquor slowly adds ethanol to pure content under constantly stirring be 70%, under 4 ℃ of conditions standing 24 hours, remove supernatant liquor, and reclaim raw sugar amine glycan after the centrifugal 10min of 8000r/min;
⑹ secondary enzymolysis: raw sugar amine glycan is dissolved in distilled water, adds 0.4% trypsinase secondary enzymolysis, pH8.0, enzymolysis time 3 hours boils the centrifugal 10min of 5000r/min after the enzyme that goes out, separation of supernatant and precipitation;
⑺ precipitate merging: all albumen precipitations merge, and make animal protein source;
⑻ CTAB in conjunction with dissociate: supernatant liquor adds the 5%CTAB(cetyl trimethylammonium bromide of 2 times of volumes), under 45 ℃, precipitation is 2 hours, the centrifugal 30min of 8000r/min, collecting precipitation, under 45 ℃, precipitation is dissolved in the CH3COONa solution of 2mol/L, the GAG-CTAB complex compound is dissociated, form dissociation solution;
⑼ alcohol precipitation, dialysis, alcohol precipitation again: above-mentioned dissociation solution adds ethanol, make the alcohol amount reach 70%, the centrifugal 10min of 8000r/min, collecting precipitation is dissolved in distilled water with precipitation, dialysis tubing dialysis 48h with the amount of damming 7000, add the Ethanol-Acetic Acid sodium reagent in dialyzate, make the alcohol amount reach 70%, press alcohol deposition method and reclaim glycosaminoglycan, with the throw out oven dry, namely make pure glycosaminoglycan.
Measure the glycosaminoglycan purity 90.56% that obtains by the A Lixin blue laws.
Example 2
⑴ raw material is processed: after the Patinopecten yessoensis tankage are cleaned, add the distilled water homogenate of 3 times of weight, make homogenate;
⑵ enzymolysis: add trypsinase and bacillus subtilis neutral proteinase in above-mentioned homogenate, the enzyme addition is 1% of tankage gross weight, and trypsinase and bacillus subtilis neutral proteinase respectively account for half, and enzymolysis is 4 hours under 40 ℃ of water bath condition; Separate and get supernatant liquor: after enzymolysis solution boils the enzyme that goes out, the centrifugal 10min of 5000r/min, separation of supernatant and precipitation;
⑶ decolouring: add 7% hydrogen peroxide decolouring in supernatant liquor;
⑷ remove protein: first transfer pH to 2.0, the centrifugal precipitation of removing with HCl; Transfer pH to 7.0, the centrifugal precipitation of removing with NaOH again;
⑸ alcohol deposition method extracts raw sugar amine glycan: supernatant liquor slowly adds ethanol to pure content under constantly stirring be 70%, under 4 ℃ of conditions standing 24 hours, remove supernatant liquor, and reclaim raw sugar amine glycan after the centrifugal 10min of 8000r/min;
⑹ secondary enzymolysis: raw sugar amine glycan is dissolved in distilled water, adds 0.4% trypsinase secondary enzymolysis, pH8.0, enzymolysis time 3 hours boils the centrifugal 10min of 5000r/min after the enzyme that goes out, separation of supernatant and precipitation;
⑺ precipitate merging: all albumen precipitations merge, and make animal protein source;
⑻ CTAB in conjunction with dissociate: supernatant liquor adds the 5%CTAB(cetyl trimethylammonium bromide of 2 times of volumes), under 45 ℃, precipitation is 2 hours, the centrifugal 30min of 8000r/min, collecting precipitation, under 45 ℃, precipitation is dissolved in the CH3COONa solution of 2mol/L, the GAG-CTAB complex compound is dissociated, form dissociation solution;
⑼ alcohol precipitation, dialysis, alcohol precipitation again: above-mentioned dissociation solution adds ethanol, make the alcohol amount reach 70%, the centrifugal 10min of 8000r/min, collecting precipitation is dissolved in distilled water with precipitation, dialysis tubing dialysis 48h with the amount of damming 7000, add the Ethanol-Acetic Acid sodium reagent in dialyzate, make the alcohol amount reach 70%, press alcohol deposition method and reclaim glycosaminoglycan, with the throw out oven dry, namely make pure glycosaminoglycan.
Measure the glycosaminoglycan purity 91.13% that obtains by the A Lixin blue laws.
Claims (4)
1. the method for a Patinopecten yessoensis tankage higher value application, is characterized in that, step is as follows:
⑴ raw material is processed: after the Patinopecten yessoensis tankage are cleaned, add distilled water homogenate, form homogenate;
⑵ enzymolysis: add trypsinase and bacillus subtilis neutral proteinase in above-mentioned homogenate, under 40 ℃ of water bath condition enzymolysis 3.5-4 hour, after enzymolysis solution boils the enzyme that goes out, the centrifugal 10min of 5000r/min, separation of supernatant and precipitation;
⑶ decolouring: add the hydrogen peroxide decolouring in supernatant liquor;
⑷ remove protein: first transfer pH to 2.0, the centrifugal precipitation of removing with HCl; Transfer pH to 7.0, the centrifugal precipitation of removing with NaOH again;
⑸ alcohol deposition method extracts raw sugar amine glycan: supernatant liquor slowly adds ethanol to pure content under constantly stirring be 70%, under 4 ℃ of conditions standing 24 hours, remove supernatant liquor, and reclaim raw sugar amine glycan after the centrifugal 10min of 8000r/min;
⑹ secondary enzymolysis: raw sugar amine glycan is dissolved in distilled water, adds 0.4% trypsinase secondary enzymolysis, pH8.0, enzymolysis time 3 hours boils the centrifugal 10min of 5000r/min after the enzyme that goes out, separation of supernatant and precipitation;
⑺ precipitate merging: all albumen precipitations merge, and make animal protein source;
⑻ CTAB in conjunction with dissociate: supernatant liquor adds the 5%CTAB(cetyl trimethylammonium bromide of 2 times of volumes), under 45 ℃, precipitation is 1-2 hour, the centrifugal 30min of 8000r/min, collecting precipitation, under 45 ℃, precipitation is dissolved in the CH3COONa solution of 2mol/L, the GAG-CTAB complex compound is dissociated, form dissociation solution;
⑼ alcohol precipitation, dialysis, alcohol precipitation again: above-mentioned dissociation solution adds ethanol, make the alcohol amount reach 70%, the centrifugal 10min of 8000r/min, collecting precipitation is dissolved in distilled water with precipitation, dialysis tubing dialysis 48h with the amount of damming 7000, add the Ethanol-Acetic Acid sodium reagent in dialyzate, make the alcohol amount reach 70%, press alcohol deposition method and reclaim glycosaminoglycan, with the throw out oven dry, namely make pure mud snail glycosaminoglycan.
2. the method for Patinopecten yessoensis tankage higher value application according to claim 1 is characterized in that: in described step ⑴ the add-on of distilled water be tankage weight 2-3 doubly.
3. the method for Patinopecten yessoensis tankage higher value application according to claim 1, it is characterized in that: in described step ⑵, the total addition level of trypsinase and bacillus subtilis neutral proteinase is the 0.8-1% of tankage gross weight, and trypsinase and bacillus subtilis neutral proteinase respectively account for half.
4. the method for Patinopecten yessoensis tankage higher value application according to claim 1, it is characterized in that: in described step ⑶, the addition of hydrogen peroxide is the 5-7% of supernatant liquor weight.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103315181A (en) * | 2013-06-27 | 2013-09-25 | 荣成宏业实业有限公司 | Shrimp and crab bait preparing from scallop zymolyte and preparing method thereof |
| CN105131143A (en) * | 2015-09-29 | 2015-12-09 | 江苏锦宇环境工程有限公司 | Preparing method of scallop viscera oligosaccharide |
| CN106117383A (en) * | 2016-06-29 | 2016-11-16 | 大连深蓝肽科技研发有限公司 | Patinopecten (Mizuhopecten) yessoensis processing byproduct is utilized to produce oligopeptide and the method for glycosaminoglycans |
| CN109053922A (en) * | 2018-07-20 | 2018-12-21 | 大连工业大学 | Scallop Viscus Polysaccharide, extracting method and its application and pharmaceutical composition |
| CN111320707A (en) * | 2020-03-31 | 2020-06-23 | 大连工业大学 | Patinopecten yessoensis skirt polysaccharide and extraction method and application thereof |
| CN113481270A (en) * | 2021-06-08 | 2021-10-08 | 中国科学院海洋研究所 | Method for extracting functional glycopeptide from scallop skirt |
| CN113502312A (en) * | 2021-06-08 | 2021-10-15 | 中国科学院海洋研究所 | Method for extracting functional glycopeptide from scallop viscera degreasing residues |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103315181A (en) * | 2013-06-27 | 2013-09-25 | 荣成宏业实业有限公司 | Shrimp and crab bait preparing from scallop zymolyte and preparing method thereof |
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| CN105131143A (en) * | 2015-09-29 | 2015-12-09 | 江苏锦宇环境工程有限公司 | Preparing method of scallop viscera oligosaccharide |
| CN106117383A (en) * | 2016-06-29 | 2016-11-16 | 大连深蓝肽科技研发有限公司 | Patinopecten (Mizuhopecten) yessoensis processing byproduct is utilized to produce oligopeptide and the method for glycosaminoglycans |
| CN109053922A (en) * | 2018-07-20 | 2018-12-21 | 大连工业大学 | Scallop Viscus Polysaccharide, extracting method and its application and pharmaceutical composition |
| CN109053922B (en) * | 2018-07-20 | 2020-12-22 | 大连工业大学 | Patinopecten yessoensis visceral polysaccharide, extraction method and application thereof, and pharmaceutical composition |
| CN111320707A (en) * | 2020-03-31 | 2020-06-23 | 大连工业大学 | Patinopecten yessoensis skirt polysaccharide and extraction method and application thereof |
| CN111320707B (en) * | 2020-03-31 | 2021-09-24 | 大连工业大学 | Patinopecten yessoensis skirt polysaccharide and extraction method and application thereof |
| CN113481270A (en) * | 2021-06-08 | 2021-10-08 | 中国科学院海洋研究所 | Method for extracting functional glycopeptide from scallop skirt |
| CN113502312A (en) * | 2021-06-08 | 2021-10-15 | 中国科学院海洋研究所 | Method for extracting functional glycopeptide from scallop viscera degreasing residues |
| CN113481270B (en) * | 2021-06-08 | 2023-07-04 | 中国科学院海洋研究所 | Method for extracting glycopeptide from scallop skirt |
| CN113502312B (en) * | 2021-06-08 | 2023-08-22 | 中国科学院海洋研究所 | Method for extracting functional glycopeptides from scallop viscera degreasing residues |
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Application publication date: 20130605 |